Biochemical and immunological properties of lactoferrin binding proteins from Moraxella (Branhamella) catarrhalis.
The Neisseriaceae can acquire iron (Fe) from lactoferrin (Lf) using host-Lf receptors on the bacterial surface. The binding proteins that are proposed to constitute the receptor have been identified by isolation with immobilized Lf. Using CopB-specific monoclonal antibodies and isogenic CopB mutants, we demonstrate that the 84 kDa protein isolated with immobilized human Lf from Moraxella catarrhalis using low stringency conditions is CopB, an 84 kDa membrane-spanning protein with similarities to other TonB-dependent outer membrane proteins. Affinity isolation of Lf receptors from a variety of M. catarrhalis strains using high stringency conditions revealed a 95 kDa protein migrating slightly faster than LbpA on SDS-PAGE in some strains. Convalescent human antisera from patients infected with M. catarrhalis reacted specifically with this protein, but not LbpA. Proteolysis experiments demonstrated that, unlike LbpA, it was rapidly degraded. The 95 kDa protein, but not LbpA, binds labelled Lf after SDS-PAGE and electroblotting, suggesting the 95 kDa protein is LbpB, the homologue of TbpB. This protein comigrates with LbpA in most strains, which may explain why it had not been previously identified.
Bonnah
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RA
Department of Microbiology and Infectious Diseases, University of Calgary, Heritage Medical Research Building, 3330-Hospital Drive, Calgary, Alberta, N.W. T2N 4N1, Canada.
Yu
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RH
Wong
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H
Schryvers
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AB
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D019286:Endopeptidase K; D006801:Humans; D007501:Iron / Q000378:metabolism; D007781:Lactoferrin / Q000378:metabolism*; D008969:Molecular Sequence Data; D001936:Moraxella catarrhalis / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development / Q000276:immunology; D011990:Receptors, Transferrin / Q000737:chemistry / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; C093197:CopB protein, Moraxella; D011990:Receptors, Transferrin; D007501:Iron; D007781:Lactoferrin; D019286:Endopeptidase K